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H Oberleithner

Publications and source records attributed to H Oberleithner.

156 records · Page 9Linked to original sources
PubMed1978

Distal site of calcium reabsorption in the rat nephron.

The site of outflux of 45Ca along the nephron were investigated using microinfusion technique in acutely thyroparathyroidectomized (TPTX), intact and TPTX Wistar rats substituted with parathyroid hormone (PTH). In all three groups 45Ca outflux occurred along the proximal tubule, the loop of Henle and along the distal tubule. After microinfusion into late distal tubules 45Ca recovery in ipsilateral urine was essentially complete for the TPTX group but was only 83 and 65% for the intact and the PTH substituted animals. Increases in microinfusion flow rate from 2--20 nl/min into early and middle distal tubules resulted in increased urinary recovery of 45Ca for all three groups. Similarily, increases in microinfusate Ca concentration from 0.2--2.0 and 5.0 mmol/l resulted in increased fractional urinary recovery of 45Ca both in the presence and absence of PTH. When 45Ca and 3H insulin containing solutions were continuously microinfused into early distal tubules for one hour periods an anticalciuric effect of PTH could be demonstrated. It is concluded that Ca, in addition to its outflux in the proximal tubule and in the loop of Henle, is reabsorbed in the distal tubule accessible to micropuncture. PTH acts anticalciuric at this latter site.

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PubMedDate not supplied

Chloride-dependence of the potency of inhibitors of the neuronal noradrenaline carrier in the rat vas deferens.

(1) Vasa deferentia obtained from reserpine-pretreated rats were exposed to 0.15 mumol l-1 3H-(-)noradrenaline (with monoamine oxidase and catechol-O-methyltransferase being inhibited) and initial rates of the neuronal 3H-noradrenaline uptake as well as IC50 values for inhibition of uptake by desipramine, cocaine or (-)metaraminol determined at various external Cl- concentrations (0-145 mmol l-1) and a fixed high Na+ concentration (145 mmol l-1). (2) When the Cl- concentration in the medium was decreased neuronal uptake fell. As far as Cl- concentrations ranging from 10 to 145 mmol l-1 are concerned, the dependence of uptake on Cl- obeyed Michaelis-Menten kinetics with an apparent Km and Vmax of 6.2 mmol l-1 and 116 pmol g-1 min-1, respectively. At Cl- concentrations below 10 mmol l-1, uptake was higher than expected from the values of Km and Vmax, and even in the nominal absence of Cl- from the medium a remainder of neuronal uptake was still detectable. Evidence is presented to show that, on incubation at Cl- concentrations below 10 mmol l-1, intracellular Cl- leaks out, so that the actual Cl- concentrations in the extracellular fluid are probably higher than in the medium. (3) The potencies of desipramine and cocaine for inhibition of neuronal uptake were markedly dependent on the Cl- concentration in the medium, but the type of Cl- -dependence differed. While the IC50 for desipramine decreased, that for cocaine increased with increasing Cl- concentration (2-145 mmol l-1).(ABSTRACT TRUNCATED AT 250 WORDS)

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PubMedDate not supplied

Extracellular detection of K+ release during migration of transformed Madin-Darby canine kidney cells.

Madin Darby canine kidney cells transformed by alkaline stress (MDCK-F cells) constitutively migrate at a rate of about 1 microm.min-1. Migration depends on the intermittent activity of a Ca2+-stimulated, 53-pS K+ channel (KCa channel) that is inhibitable by charybdotoxin. In the present study we examined whether this intermittent KCa channel activity results in a significant K+ loss across the plasma membrane. K+ efflux from MDCK-F cells should result in a transient increase of extracellular K+ ([K+]e) in the close vicinity of a migrating cell. However, due to the rapid diffusion of K+ ions into the virtually infinite extracellular space, such a transient increase in [K+]e was too small to be detected by conventional K+-selective electrodes. Therefore, we developed a "shielded ion-sensitive microelectrode" (SIM) that limited diffusion to a small compartment, formed by a shielding pipette which surrounded the tip of the K+-sensitive microelectrode. The SIM improved the signal to noise ratio by a factor of at least three, thus transient increases of [K+]e in the vicinity of MDCK-F cells became detectable. They occurred at a rate of 1.3 min-1. The cell releases 40 fmol K+ during each burst of intermittent KCa channel activity, which corresponds to about 15% of the total cellular K+ content. Since transmembrane K+ loss must be accompanied by anion loss and therefore leads to a decrease of cell volume, these findings support the hypothesis that intermittent volume changes are a prerequisite for the migration of MDCK-F cells.

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PubMedDate not supplied

Aldosterone and nuclear signaling in kidney.

Initiation of transcription is an early step in steroid hormone action. We investigated by means of atomic force microscopy (AFM) and fluorescence imaging the role of nuclear pore complexes (NPCs) in mediating signal transduction of the mineralocorticoid hormone aldosterone from the extracellular space into the cell nucleus. With AFM, we imaged single NPCs of isolated nuclear envelopes under native conditions. We observed that individual NPCs contract in response to a Ca2+ signal, which is known to occur in seconds after aldosterone exposure. In living kidney cells in culture (MDCK cells), aldosterone led within seconds to the contraction of the whole nucleus measured by DNA-fluorescence imaging. Nuclear contraction was elicited at similar time scale and to similar extent by bradykinin, a peptide hormone known to mobilize Ca2+ from internal stores, and by ionomycin, a Ca2+ ionophore known to directly increase intracellular Ca2+. Nuclear contraction is explained by the individual contraction of calcium-sensitive NPCs that occur in high density in the nuclear envelope. We present a model in which nuclear pore complexes play a key role as barrier molecules of high plasticity in the control of aldosterone-induced gene expression.

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PubMedDate not supplied

Aldosterone remodels human endothelium.

AIM: In response to aldosterone endothelial cells swell and stiffen. Although amiloride-sensitive sodium and water uptake is known to be involved, the underlying mechanisms are yet unclear. We tested the hypothesis whether the intracellular accumulation of water or organic matter is responsible for the structural and functional alterations. METHODS: Atomic force microscopy was used as an imaging tool and a mechanical nanosensor. Cell water, organic cell matter and cell pressure was measured at single cell level in human umbilical vein endothelial cells (HUVEC). Furthermore, we tested by means of a miniature perfusion chamber in vitro the physical robustness to blood flow of the aldosterone-treated endothelium. RESULTS: In response to a three-day treatment with 1 nM aldosterone HUVEC swell. To our surprise, cell water decreased from 82+/-6% to 71+/-5% while intracellular organic matter increased from 18+/-1.8% to 29+/-3.0%. These changes were paralleled by a rise in cell pressure of 114%, measured in living HUVEC in vitro. Blood flow across the endothelium was found significantly altered after aldosterone treatment. Imaging the endothelial monolayer after blood perfusion disclosed large gaps between cells treated with aldosterone. The mineralocorticoid receptor blockers, spironolactone and eplerenone could prevent the aldosterone actions. CONCLUSION: Mild aldosteronism causes intracellular accumulation of organic matter at the cost of cell water. This makes endothelium stiff and vulnerable to shear stress. The measurements could explain clinical observations that high blood pressure combined with high plasma aldosterone concentration may damage the endothelium of blood vessels.

Aldosterone↗
PubMedDate not supplied

Carbonic anhydrase activity in Madin Darby canine kidney cells. Evidence for intercalated cell properties.

Madin Darby canine kidney (MDCK) renal epithelial cell cultures have been investigated with respect to their potency to express carbonic anhydrase activity using histochemical methods. Acetazolamide inhibitable carbonic anhydrase activity could be detected in the cytoplasmic compartment as well as in the apical membrane of cells when grown on solid culture supports. Cells forming domes in MDCK monolayers exhibit the highest histochemically detectable enzyme activity. The attempt to subculture clonal cell lines from MDCK monolayer cultures resulted in the establishment of 5 clones, slightly different with respect to size and shape of cells and their potency to form domes. Scanning electron microscopy ensured the identification of one clone (1A4), which distinctly differed from the others with respect to the apical membrane architecture. Co-localization of peanut agglutinin and carbonic anhydrase activity at the plasma membrane always revealed a combined occurrence of enzyme reactivity and lectin binding in the apical membrane domain. Both, lectin binding and carbonic anhydrase activity were distinctly more intense in plasma membrane regions equipped with microvilli. From the results it is concluded that MDCK cells in tissue culture retained properties of intercalated cells of the nephron collecting duct segment.

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